Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1997-06-02 to 1997-07-18
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted July 17, 1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
96/54/EC
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Sodium ascorbyl phosphate
- Physical state: Powder, beige
- Analytical purity: 85.4 g/100 g (HPLC)
- Date of Manufacturing: 1997-05-02
- Lot/batch No.: 28600/37-9
- Stability under test conditions: The stability of the test substance in aqua bidest. for a time period of 96 hours was confirmed by analysis
- Storage condition of test material: Room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River GmbH - Wiga, Kisslegg, Germany
- Age at study initiation: Young adult animals
- Weight at study initiation: 320 - 394 g
- Housing: 5 animals per cage (Makrolon, type IV), Bedding: Granulat Type 3/4 (staubfrei) SSNIFF
- Diet: Kliba Labordiät 341 (Kaninchen-Meerschweinchen-Haltungsdiät) ad libitum
- Water: Water ad libitum (tap water; about 2 g of ascorbic acid per 10 L water was added to the drinking water twice a week)
- Acclimation period: 7 days before the beginning of the study in the laboratory for dermal toxicity


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 °C
- Humidity (%): 30-70 %
- Photoperiod (hrs dark / hrs light): 12 h light (6.00 a.m. - 6.00 p.m.), 12 h darkness (6.00 p.m. - 6.00 a.m.)


IN-LIFE DATES: From: 1997-06-10 To: 1997-07-11

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
other: intradermal and percutaneous occlusive
Vehicle:
water
Concentration / amount:
50%
Challengeopen allclose all
Route:
other: intradermal and percutaneous occlusive
Vehicle:
water
Concentration / amount:
50%
No. of animals per dose:
Pretest: 4
Main test: 10 (control), 30 (test group)
Details on study design:
RANGE FINDING TESTS:
Amount applied:
2 x 2 cm filter paper strips containing the test substance (5 %, 10 %, 25 %, 50 %) formulation were applied to the skin of the flanks under an occlusive dressing. The filter paper strip was soaked in the test substance formulation. The dressing consisted of rubberized linen patches (4 x 4 cm from Russka), patches of Idealbinde (5 x 5 cm from Pfälzische Verbandstoff-Fabrik) and Fixomull®D Stretch (adhesive fleece) from Beiersdorf AG.
Exposure period:
The test substance was applied 2 times for 24 hours within a period of 96 hours in order to detect nonspecific phenomena that are not caused by a sensitization reaction but could possibly be attributed to a shift in the irritation threshold.
Site of application:
- flank, respectively on the same area
Number of test animals:
- 4 per test concentration
Readings:
- about 24 and 48 h after the beginning of application
Assessment of skin findings (according to Draize, J.H. (1959) : Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. The Association of Food and Drug Officials of the United States Austin, Texas)



MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 6 intradermal injections in groups of two per animal
- Exposure period: 48 h
- Test groups:
A) front row: 2 injections each of 0.1 mL Freund's adjuvant without test substance emulsified with 0.9% aqueous NaCL-solution in a ratio of 1 :1
B) middle row: 2 injections each of 0.1 mL of the test substance formulation
C) back row: 2 injections each of 0.1 mL Freund's adjuvant / 0.9% aqueous NaCl-solution (1 : 1) with test substance
- Control group:
The animals were given the same injections (A, B,C) but without test substance, only with the formulating agent.
- Site: shoulder
- Duration: 48 h
- Concentrations: 2 x 4 cm filter paper strips containing the test substance (50%) formulation were applied to the skin of the shoulder under an occlusive dressing. The filter paper strip was soaked in the test substance formulation. The dressing consisted of rubberized linen patches (4 x 6 cm from Russka) and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG. The control animals were not treated since the distilled water used as formulating agent was not expected to influence the result of the study.


B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: Day 14, Day 21
- Exposure period: 24 h
- Test and Control groups:
1st challenge:
- The test group and control group 1 were treated with the test substance formulation (control group 2 remained untreated).
2nd challenge:
- The test group and control groups 1 and 2 were treated with the test substance formulation.
- Site: intact flank
- Concentrations: 2 x 2 cm filter paper strips containing the test substance (50%) formulation were applied to the skin of the flank under an occlusive dressing. The filter paper strip was soaked in the test substance formulation. The dressing consisted of rubberized linen patches (4 x 4 cm from Russka), patches of Idealbinde (5 x 5 cm from Pfälzische Verbandstoff-Fabrik) and Fixomull®D Stretch (adhesive fleece) from Beiersdorf AG.
- Evaluation (hr after challenge): 24 and 48 h after the removal of the patch
Challenge controls:
1st challenge:
- The control group 1 was treated with the test substance formulation (control group 2 remained untreated).
2nd challenge:
- The control groups 1 and 2 were treated with the test substance formulation.
Positive control substance(s):
yes
Remarks:
A positive control (reliability check) with Alpha-Hexylcinnamaldehyde is not included in this study. However, a separate study is performed twice a year in the laboratory.

Results and discussion

Positive control results:
A positive control (reliability check) with a known sensitizer is not included in this study. However, a separate study is performed twice a year in the laboratory.
The positive control with Alpha-Hexylcinnamaldehyde techn. 85% showed that the test system was able to detect sensitizing compounds under the laboratory conditions chosen.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
other: first challenge
Hours after challenge:
24
Group:
test group
Dose level:
50 %
No. with + reactions:
2
Total no. in group:
23
Clinical observations:
7 test group animals died 8, 9 or 10 days after the beginning of the study. Macroscopic examination revealed that animals suffered from pneumonia (not test substance treatment related). Well-defined, spotted erythema was observed in 2 test group animals.
Remarks on result:
other: see Remark
Remarks:
Reading: other: first challenge. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 2.0. Total no. in groups: 23.0. Clinical observations: 7 test group animals died 8, 9 or 10 days after the beginning of the study. Macroscopic examination revealed that animals suffered from pneumonia (not test substance treatment related). Well-defined, spotted erythema was observed in 2 test group animals..
Reading:
other: first challenge
Hours after challenge:
48
Group:
test group
Dose level:
50%
No. with + reactions:
3
Total no. in group:
23
Clinical observations:
3 test group animals showed very slight, spotted erythema, 2 out of these animals additionally exhibited scaling. In 1 test group animal well defined, spotted erythema and scaling were noted.
Remarks on result:
other: see Remark
Remarks:
Reading: other: first challenge. . Hours after challenge: 48.0. Group: test group. Dose level: 50%. No with. + reactions: 3.0. Total no. in groups: 23.0. Clinical observations: 3 test group animals showed very slight, spotted erythema, 2 out of these animals additionally exhibited scaling. In 1 test group animal well defined, spotted erythema and scaling were noted..
Reading:
other: first challenge
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none (control group 1)
Remarks on result:
other: Reading: other: first challenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none (control group 1).
Reading:
other: first challenge
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none (control group 1)
Remarks on result:
other: Reading: other: first challenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none (control group 1).
Reading:
other: second challenge
Hours after challenge:
24
Group:
test group
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
23
Remarks on result:
other: Reading: other: second challenge. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 23.0.
Reading:
other: second challenge
Hours after challenge:
48
Group:
test group
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
23
Remarks on result:
other: Reading: other: second challenge. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 23.0.
Reading:
other: second challenge
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none (control group 1)
Remarks on result:
other: Reading: other: second challenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none (control group 1).
Reading:
other: second challenge
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none (control group 1)
Remarks on result:
other: Reading: other: second challenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none (control group 1).
Reading:
other: second challenge
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none (control group 2)
Remarks on result:
other: Reading: other: second challenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none (control group 2).
Reading:
other: second challenge
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none (control group 2)
Remarks on result:
other: Reading: other: second challenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none (control group 2).

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Sodium ascorbyl phosphate is not a skin sensitiser.